Coding

Part:BBa_K4275005

Designed by: Wang Sze Ting   Group: iGEM22_GreatBay_SCIE   (2022-09-29)
Revision as of 22:54, 11 October 2022 by Jerryatcg (Talk | contribs)


MtCDH-t

MtCDH-t, fused with a dockerin, is a cellobiose dehydrogenase, one of the five cellulose-related enzymes fixed on type I cohesin, and thus on the scaffold composed of CipA2B9C and OlpB. Its reductive nature enables it to act as an electron donor to the “cellulase booster”, TaLPMO-t, another cellulose-related enzyme on the scaffold which boosts the efficiency of crystalline cellulose degradation. By fusing a type 1 dockerin through a CBM and a 36-bp glycine-rich linker at the C terminal of MtCDH, the free fungal reductase is converted into the cellulosomal mode. Synergizing with the other four cellulose-related enzymes and cellulose binding modules, MtCDH-t is an important contributor to the enhanced efficiency of cellulose degradation. This is a part in a part collection where we enable efficient degradation of cellulose in textile waste.

GreatBay SCIE--3D MtCDH-t.png

Usage and Biology

MtCDH-t, fused with a dockerin, is a cellobiose dehydrogenase that enhances cellulose degradation by coupling the oxidation of cellobiose to the reductive activation of polysaccharide monooxygenases (PMO) that catalyze the insertion of oxygen into C−H bonds adjacent to the glycosidic linkage. MtCDH has a heme domain at the N-terminal and a flavin domain at the C-terminal. The flavin domain is the site of oxidation of cellobiose, and subsequently, electrons are transferred to the heme domain. The reduced heme domain reduced the PMOs. MtCDH-t is fused with a type 1 dockerin through a CBM and a 36-bp glycine-rich linker, thus can bind to type 1 cohesin of the scaffold, immobilizing the enzyme.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2706
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1507
    Illegal AgeI site found at 358
    Illegal AgeI site found at 745
    Illegal AgeI site found at 871
    Illegal AgeI site found at 1420
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None